Industrial Crops and Products 80 (2016) 93–100

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Industrial Crops and Products

journal homepage: www.elsevier.com/locate/indcrop

The green generation of sunscreens: Using coffee industrial

sub-products
J. Marto a , L.F. Gouveia a , B.G. Chiari b , A. Paiva c , V. Isaac b , P. Pinto a , P. Simões c ,
A.J. Almeida a , H.M. Ribeiro a,∗
a
Research Institute for Medicines and Pharmaceutical Sciences (iMed.UL), Faculty of Pharmacy, University of Lisbon, Portugal
b
Faculdade de Ciências Farmacêuticas, UNESP – Univ Estadual Paulista, DFM – Laboratório de Cosmetologia – LaCos, São Paulo, Brazil
c
LAQV-REQUIMTE, Departamento de Química,Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal

a r t i c l e i n f o a b s t r a c t

Article history: Spent coffee grounds and green coffee defective beans, which are industrial sub-products of coffee pro-
Received 6 July 2015 cessing, have a potential use for cosmetic applications, due to their safety and high content in lipids that
Received in revised form 4 November 2015 present interesting physicochemical properties. Sunscreen formulations in the form of water-in-oil emul-
Accepted 9 November 2015
sions might be a suitable application for these sub-products because providing a higher sun protection
factor (SPF) for the same concentration of sunscreen activities than oil-in-water emulsions.
Keywords:
The purpose of this work was to assess the biological effects of using the oil fraction of spent coffee
Spent coffee oil
grounds extracted with supercritical CO2 and green coffee oil in the development of new generation of
Green coffee oil
Supercritical fluid extraction
sunscreens with improved sun protection performance. The oil fractions were used to prepare w/o sun-
Water resistant screens involving a cold emulsification process, with purified water as disperse aqueous phase and TiO2
Pickering emulsion sunscreen and ZnO particles as stabilizers. The sunscreens were characterized in terms of mechanical, rheological
and skin adhesion properties. In addition, the in vitro and in vivo biological properties of the formulations
were evaluated, including safety and sunscreen water resistance tests.
The use of two types of solid particles proved to be useful in the developed formulations, ensuring
a high SPF with UVB/A protection, conferred by TiO2 and ZnO, respectively. Moreover, the emulsion
containing 35% w/w of the spent coffee grounds oil fraction presented promising characteristics in the
improvement of water performance with a broad spectrum sun protection when compared to an emulsion
containing 35% w/w of green coffee oil which improved the SPF in physical sunscreens. The formulations
are industrial-scalable and suitable for topical use according to the rheological, mechanical and safety
assessment.
The use of spent coffee oil in cosmetic industry seems to be a suitable approach for the valorisation of
waste from the coffee industry and presents promising characteristics in the improvement of sunscreen
performance.
© 2015 Elsevier B.V. All rights reserved.

1. Introduction Avoiding sun exposure, covering the skin or applying sunscreens

The efficacy of sunscreen products has been recognized as an
important public health issue and is usually expressed by the sun
protection factor (SPF), which is calculated as the ratio between
the UV energy required to produce a minimal erythema dose of
protected and unprotected skin (Dutra et al., 2004; Ribeiro et al.,
2013).
∗ Corresponding author at: Research Institute for Medicines (iMed.ULisboa), Fac-
ulty of Pharmacy, Universidade de Lisboa Av. Professor Gama Pinto, 1649-003 Lisboa,
Portugal. Fax: +351 217946470.
E-mail address: hribeiro@campus.ul.pt (H.M. Ribeiro).
with a high SPF are the main strategies strongly recommended to
prevent UV-induced cell damage. The UV filters can be divided in
two groups: (a) chemical filters which absorb UV radiation (UVR);
and (b) physical filters, such as, titanium dioxide (TiO2 ) which
reflect UVR (Ascenso et al., 2014).
Sunscreens are normally based on synthetic chemicals with high
capacity to absorb sun light at the region of UVB (320–290 nm) and
UVA (400–320 nm) spectrum. Several synthetic UV filter molecules
(e.g. benzophenones, anthranilates, PABA derivatives, salicylates,
cinnamates and camphor derivatives) are available as photopro-
tective agents, but due to their harmful effects they are becoming
less popular. The main problem of the chemical sunscreen agents

http://dx.doi.org/10.1016/j.indcrop.2015.11.033
0926-6690/© 2015 Elsevier B.V. All rights reserved.
94 J. Marto et al. / Industrial Crops and Products 80 (2016) 93–100
are the photoirritation, photosensitization and contact dermati-
tis. Reducing the concentration of such chemicals in cosmetics is a
strategy to improve their quality, without affecting their properties
(Serpone et al., 2007).
In recent years, the use of natural agents has been attracting sig-
nificant attention, due to their safety, multiple biological actions
on the skin and cost effectiveness. Consequently, an oily vehicle
with antioxidant activity could be an excellent approach for its
important role on the product efficacy by improving the prod-
ucts’ photoprotective activity (Calixto et al., 2011; Couto et al.,
2009). In addition to bioactivity, natural products are, in general,
not harmful for humans, not expensive, suitable to be used in a wide
range of applications, and are obtained from renewable sources.
In this context, green coffee oil and spent coffee grounds oil have
arisen as potential candidates to replace synthetic chemicals in sun-
screens since they are a rich source of antioxidants and polyphenols
(Ribeiro et al., 2013). Phenolic compounds are excellent candidates
for the prevention of the harmful effects of UV radiation on the
skin. More specifically, flavonoids have photoprotection potential
due to their UV absorbing capacity, ability to act as antioxidants
and anti-inflammatory, and immunomodulatory agents (Saewan
and Jimtaisong, 2013).
Coffee consumption is growing worldwide, being extremely
embedded in the cultural habits of many countries, so the total
import and export of coffee has been increasing. Spent coffee
grounds (SCG), which are the residue obtained after the treatment
of coffee with hot water or steam for extracting flavour substances,
can be used for industrial applications such as high quality biodiesel
production (Couto et al., 2009). Nevertheless, due to their high con-
tent in lipids, particularly fatty acids, SCG might also find a suitable
application in cosmetic products where these lipid compounds can
be used as valuable excipients. Therefore, the lipid fraction of SCG
extracted with supercritical CO2 can be used in the development
of new, improved sunscreens. On average, a fifth of the Brazilian
coffee production consists of defective beans, and several studies
have been developed in order to find an alternative use for sub-
product, including the cosmetic application of the extracted oil
(Preedy, 2014; Ribeiro et al., 2013).
Titanium dioxide has been incorporated in sunscreen formula-
tions for more than 25 years, being regarded as safe and effective,
thus bringing together two of the most desirable features in cos-
metic market (Renner, 2009). It is especially preferred by people
with a high propensity for skin irritation, such as patients under-
going oncological chemotherapy. Moreover, TiO2 particles are a
UV-B filter, suitable for developing physical sunscreens combining
both increasing stability and high SPF properties (Wang and Tooley,
2011). The introduction of ZnO ensures an adequate protection in
the range of UVA. Due to its multifunctional nature, ZnO particles
have been shown to be effective as antibacterial and antifungal
agent (Singh et al., 2012; Smijs and Pavel, 2011).
On the other hand, TiO2 particles can be also used as solid parti-
cles for stabilization of surfactant-free emulsions stabilized by solid
particles (i.e. Pickering emulsions). This type of emulsions have
important advantages over the classical surfactant-based emul-
sions, such as higher resistance to coalescence due to an improved
stability, and a higher tolerability (Laredj-Bourezg et al., 2012). The
stabilization of emulsion droplets takes place by means of adsorp-
tion of solid particles at the surface of emulsion droplets. It can
be assumed that a stable water-in-oil (w/o) Pickering emulsion is
a function of particles concentration, pH and ionic strength. This
adsorption mechanism is quite different compared to surfactants
since partial wetting of the solid particles surface by water and oil is
the reason of the strong anchoring of these particles at the water–oil
interface. Few fully natural and biocompatible materials are avail-
able for the effective stabilization of these emulsions since severe
requirements must be simultaneously met, including insolubility in
both fluid phases and intermediate wettability (Folter et al., 2012).
The purpose of this work was to develop and characterize a w/o
emulsion stabilized by physical sunscreens containing 35% of the
lipid fraction of spent coffee grounds extracted with supercriti-
cal CO2 and green coffee oil obtained from defective beans with
improved sunscreen performance. Sunscreen formulations might
be a suitable application for these types of sub-products because
w/o emulsions are water resistant and provide greater efficacy (a
higher SPF) for the same concentration of sunscreen actives than
their o/w counterparts (Couteau et al., 2012).
2. Material and methods
2.1. Materials
Spent coffee grounds oil (SCO) was supplied by LAQV-
REQUIMTE – Departamento de Química, Faculdade de Ciências e
Tecnologia (Caparica, Portugal). Green coffee oil (GCO) was sup-
plied by Cooxupé – Cooperativa de Cafeicultores de Gauxupé,
(Minas Gerais, Brazil). Triethoxycaprylylsilane titanium dioxide
(mTiO2 ) (Unipure White LC 987) was a gift from Sensient (Milwau-
kee, USA). The starch used was aluminum starch octenylsuccinate
(ASt) (DryFlo® Plus) obtained from AkzoNobel (Amsterdam,
Netherlands). Zinc oxide (ZnO) (Tego® Sun Z 500) was obtained
from Evonik Industries AG (Essen, Germany). Purified water was
obtained by inverse osmosis (Millipore, Elix® 3).
2.2. Methods
2.2.1. Characterisation of the formulation ingredients
2.2.1.1. Wettability measurements. Contact angles of water, green
coffee oil and spent coffee oil on ZnO, mTiO2 and ASt in air atmo-
sphere were measured at room temperature by using ConAnXL—a
Microsoft Excel based workbook and add-in software (freely avail-
able upon request) as described in detail elsewhere (Marto et al.,
2015). All measurements were performed in triplicate.
2.2.1.2. Particle size distribution. Particle size distribution was
determined using a Malvern Mastersizer 2000 (Malvern Instru-
ments, UK), coupled with a Hydro S accessory. Data were expressed
in terms of relative distribution of volume of particles in the range
of size classes, and given as diameter values corresponding to per-
centiles of 10, 50 and 90. The Span value is a useful parameter to
characterize the particle size distribution broadness.
2.2.1.3. Natural oils.
2.2.1.3.1. Oil extraction. SCG oil was obtained by supercriti-
cal CO2 extraction as described elsewhere (Ribeiro et al., 2013).
Supercritical fluid extraction can be an environmentally friendly
alternative to traditional organic solvent extraction processes
whereby extraction/separate recovery of oil and bioactive com-
pounds from agro-industrial residues can be done without their
degradation (Brunner, 2013). Mass transport is highly facilitated
owing to favourable transport properties (high mass and thermal
diffusivities coupled with low viscosities) and the solvation capac-
ity of the supercritical fluid is tuneable by changing the operating
conditions of pressure and temperature. The most commonly used
supercritical fluid, carbon dioxide, is non-toxic, non-flammable,
non-corrosive, relatively inert from a chemical point of view and
environmentally friendly. Its relatively low critical temperature
(304.3 K) allows extraction of thermolabile substances without
degradation.
The supercritical CO2 extraction of SCG oil was done in a high
pressure extraction pilot unit. SCG were first dried in an oven at
 J. Marto et al. / Industrial Crops and Products 80 (2016) 93–100
Table 1
Qualitative and quantitative composition of the sunscreen formulations.
Ingredients Quantitative composition (%, w/w)
GCO sunscreen SCO sunscreen
Phase A
Green coffee oil 35 –
Spent coffee oil – 35
Phase B
Triethoxycaprylylsilane titanium dioxide 20 20
Zinc oxide 15 15
Aluminum starch octenylsuccinate 5 5
Phase C
Purified water 25 25
378 K with air circulation until a moisture value of about 1%. Super-
critical CO2 extractions were carried out at 55 ◦ C and 250 bar using
0.5 kg of dry SCG per batch with an average CO2 flowrate of 15 kg/h
for ca. 1 h of extraction.
GCO was obtained by cold pressed extraction. Briefly, green cof-
fee beans were pressed and come back to be pressed again for a
further three times and then the oil is bottled with nitrogen.
2.2.1.3.2. SPF measurement. GCO and SCO were accurately
weighed (0.25 g), diluted with ethanol, followed by ultrasonication
for 5 min and filtered through filter paper (WhatmanTM 42). The
absorption spectra of samples solution were obtained in the range
of 290–320 nm (Hitachi U-2001, USA) every 5 nm, using a standard
1 cm quartz cell, and ethanol as the blank. Triplicates were made,
followed by the application of the Mansur equation (Eq. (1)).
SPFspectrophotometric = CF × 320
290 EE () × I() × Abs()
where EE () is the erythemal effect spectrum; I () is the solar
intensity spectrum; Abs () is the absorbance of sunscreen product;
CF is the correction factor (=10) (Mansur et al., 1986). The values of
EE × I are constants determined by Sayre et al. (1979).
2.2.2. Sunscreen formulations—green coffee oil sunscreen (GCO
sunscreen) and spent coffee oil sunscreen (SCO sunscreen)
Two formulations were developed (Table 1) based on their
macroscopic appearance, physical stability and SPF value. The
disperse aqueous phase was composed of purified water, the con-
tinuous oil phase consisted of GCO or SCO. Solid particles (mTiO2 ,
ZnO and ASt) were firstly dispersed in the oil phase. The oil and
aqueous phases were then mixed using a high-speed homogenizer
(UltraTurrax® , IKA-Werke GmbH & Co., KG, Germany) at room tem-
perature (cold process).
2.2.3. Characterization of the sunscreen formulations
2.2.3.1. Droplet size distribution. The emulsions were examined
1 day after preparation, using an optical microscope (Olympus
CX40, Japan) equipped with a video camera. The droplet size was
determined using the image analysis software Olympus Stream
Essentials® . The size data was expressed in terms of relative size
distribution of particles (BS, 1993).
2.2.3.2. Structural analysis of the sunscreen formulations.
2.2.3.2.1. Rheology studies. Shear rate vs shear stress measure-
ments were performed at 25 ◦ C using a HAAKE RS-1 Rheometer,
equipped with automatic gap setting (Thermo ScientificTM ,
Waltham, USA). Rotational viscosity was determined using a
C35 mm cone geometry, with an angle of 2◦ . Dynamic or shear
viscosity measurements were carried out by rotational shear exper-
iments, between 1 and 1000 Pa. Oscillation sweep tests were
performed at frequencies ranging between 0.1 and 100 Hz. The
creep and recovery test was carried out with a shear stress of 1 Pa
for emulsions, allowing 360 s for creep and 360 s for relaxation. All
95
tests were performed on samples of about 1 g, at 25 ± 0.5 ◦ C. All
experiments were performed in triplicate using new samples for
each measurement.
2.2.3.2.2. Texture profile analysis (TPA). A Texture Analyzer
TA.XT Plus with Exponent 3.0.5.0 software (Stable Micro Systems
Ltd., Godalming, UK) was used to examine hardness, elasticity,
compressibility, adhesiveness and cohesiveness of the emulsion.
A probe (P/10, 10 mm Delrin), which was twice depressed into
the sample at a defined rate (5 mm/s) to a desired depth (15 mm),
allowing 15 s of delay between consecutive compressions. Six repli-
cates were performed at 25 ◦ C for each formulation.
2.2.3.3. In vitro SPF. The SPF was assessed using the Optometrics
SPF-290S Analyzer (Optometrics Corporation, Essex, UK). The sam-
ples were prepared by spreading 110 mg of each formulation over
a Transpore® tape (70.7 × 70.7 mm) to obtain a film of 2 mg/cm2 ,
as specified by the European Regulation (EC, 2009). Each sample
was exposed to a xenon arc solar simulator, and the analyser per-
formed scans in 6 different spots on the Transpore® tape substrate.
Each scan takes a transmittance (T) measurement every 2 nm from a
wavelength ranging from 290 to 400 nm. The Monochromatic Pro-
tection Factor (MPF) was determined for the selected wavelengths
using Eq. (2). The SPF value was calculated using Eq. (3).
1
MPF = (2)
T
400
290
E.B
SPF = 400 E.B (3)
290 MPF
(1)
where (E) is the spectral irradiance of terrestrial sunlight under
controlled conditions and (B) is the erythema effectiveness (Kale
et al., 2010).
2.2.3.4. In vitro sun product water resistance. The water resistance
of developed sunscreens was measured using an improved in vitro
bath system. An amount of 2 mg/cm2 of sunscreen formulation
was dispensed onto the plate, and carefully applied with a rubber-
gloved finger. After drying for 15 min, the SPF of each sample was
determined using the SPF 290 analyzer (Optometrics SPF-290S Ana-
lyzer). The samples were immersed in the in vitro bath system
(29 ± 2 ◦ C) and washed away by the water flow (150 rpm) during
20 min. The samples were allowed to air dry for 15 min and SPF
was measured again. The samples were immersed once more and
washed during 20 min. The samples were allowed to air dry for
15 min and SPF was measured to calculate the water resistance
retention (%WRR) of the sunscreens, as defined by Eq. (4).
SPFwet
%WRR = × 100 (4)
SPFdry
where SPFdry and SPFwet are the SPFs before and after water immer-
sion, respectively (Ahn et al., 2008; COLIPA, 2005).
2.2.3.5. Skin adhesion properties. Skin adhesion measurements
were performed using Texture Analyzer TA.XT Plus (Stable Micro
Systems Ltd., Surrey, UK) equipped with a load cell of 5 kg, cylin-
der probe of 10 mm (P/10, 10 mm Delrin) and a measuring system
A/MUC (skin adhesion test rig), which holds the human skin. An
amount of 2 mg/cm2 of each sample was applied onto the skin.
The probe was immersed in an in vitro bath system (29 ± 2 ◦ C) and
washed away by the flow of water (150 rpm) during 40 min, in
order to simulate the water resistance method for sun protection
products (COLIPA, 2005). The area under the curve (AUC) was calcu-
lated from the force–distance plot as the work of skin adhesion per
square centimetre. The formulation given below was used to calcu-
96 J. Marto et al. / Industrial Crops and Products 80 (2016) 93–100

late the work of skin adhesion per square centimetre (mJ/cm2 —Eq. Table 2
Contact angle of water, GCO and SCO on mTiO2 , ZnO and ASt (mean ± SD, n = 3).
(7)). Each experiment was carried out in triplicate.
Contact angle (◦ )
  AUC
Workofskinadhesion mJ/cm2 = (7) Samples Water Green coffee oil Spent coffee oil
r 2
mTiO2 106.5 ± 0.7 60.1 ± 1.9 9.7 ± 3.3
whereby r2 is the surface area of the human skin that is in contact ZnO 100.2 ± 2.6 23.4 ± 0.1 13.1 ± 2.5
with the sunscreen formulations. ASt 109.0 ± 0.4 10.6 ± 2.3 13.3 ± 4.0

2.2.3.6. Human repeat insult patch test (HRIPT). A safety evalua-
tion study was performed on emulsions, using the Marzulli and
Maibach (1976) HRIPT protocol. In brief, the product was applied
on the back of 51 healthy volunteers that gave their prior informed
written consent. For the induction period, a series of nine patches
(Finn Chamber standard) were performed over a period of 3 weeks.
Reactions after patching were scored according to International
Contact Dermatitis Research Group (Fregert and Bandmann, 1975).
A 2 weeks rest period was maintained without application of the
test material. During the challenge period, new patches were pre-
pared and fixed in the same manner as in the induction period, but
also on the right side of the back (i.e. a virgin site).
The protocol was approved by the local Ethical Committee and
respected the Helsinki Declaration and the AFSSAPS regulations on
performed HRIPT studies on cosmetic products. The study was con-
ducted under the supervision of a dermatologist who participated
in the evaluation of irritation/allergic reactions to the emulsions.
2.2.3.7. In vivo sun product water resistance. The water resistance of
sunscreens was tested on 3 subjects (Fitzpatrick skin type II). Pan-
ellists cleanse their forearms using a mild cleanser and leave them
to air dry for 30 min before starting the test. Initial cross-polarized
images were taken after the sunscreens application (2 mg/cm2 ) on
the inner forearm (4 cm2 ).
The amount of each sunscreen formulation left before and after
water bath immersion was quantified via cross polarized imaging
by means of the Visia® CA (Canfield Scientific, Faitfield, NJ). Panel-
lists immerse their forearms into a water bath system (29 ± 2 ◦ C)
and washed away by the flow of water (150 rpm) during 40 min
(Ahn et al., 2008).
The RGB colour space of the raw bitmap images was converted
to relative luminance using ImageJ® . From these images, average L
changes for each sunscreen area were obtained from histograms.
Skin whiteness was defined as the change in L value before and after
water immersion, and the percentage of water resistance retention
(%WRR) of the sunscreens was determined according to Eq. (8).
Lwashedprotector − Lskin
%WRR =
Lprotector − Lskin
2.2.4. Statistical analysis
The data are expressed as mean and standard devia-
tion (mean ± SD) of experiments. Statistical evaluation of data
was performed using one-way analysis of variance (ANOVA).
Tukey–Kramer multiple comparison test (GraphPad PRISM 5 soft-
ware, USA) was used to assess the significance of the difference
between the groups (p < 0.05).
3. Results and discussion
3.1. Characterisation of the formulation ingredients
the particle makes with it, which will determine the type of emul-
sion. Generally, if the water contact angle is <90◦ , the solid surface
is considered hydrophilic and if the water contact angle is >90◦ , the
solid surface is considered hydrophobic. Concerning the emulsions
stabilized by solid particles, if the contact angle, measured through
the aqueous phase, is <90◦ the emulsion will be o/w and, by con-
trast, if the contact angle is >90◦ the emulsion will be w/o (Marku
et al., 2012). In this study, mTiO2 , ZnO and ASt will stabilized more
effectively w/o emulsions. All solid particles have a contact angle
with water >90◦ , due to hydrophobic coatings, and simultaneously,
a contact angle with Green coffee oil (GCO) and Spent coffee oil
(SCO) <90◦ (Table 2). Thus, by combining these three particles it is
possible to obtain a stable w/o emulsion.
3.1.2. Particle size distribution
Particle size distributions of mTiO2 , ZnO and ASt showed that all
particles were larger than 100 nm (Table 3), thus complying with
the international “green” standards, whereby nanomaterials must
be avoided (COSMOS, 2013).
3.1.3. Natural oils
3.1.3.1. SPF measurement. Several studies have been developed in
order to find an alternative use for these defective coffee beans
or spent coffee ground, involving the characterization of their
lipid fractions. The GCO was extracted using mechanical pressing,
whereas the lipid fraction of SCG was extracted with supercritical
CO2 , an environmentally friendly solvent that allows the extraction
and recovery of the oil at such conditions that no degradation of the
lipid composition oil occurs while avoiding the use of hazardous
organic solvents (Ribeiro et al., 2013).
The GCO and SCO have potential application in health care
products, including sunscreen formulations (Chiari et al., 2014).
In particular, due to the richness of polyphenols and flavonoids,
the pure GCO shows a SPF higher than 5, improving SPF and,
consequently, decreasing the concentration of chemical and/or
physical sunscreens in such formulations (Wagemaker et al., 2011).
In this study, GCO presented an SPF value of 5.03 ± 0.23 while
(8) SCO presented only 1.57 ± 0.07, which is in line with the fact that
the coffee roasting process diminishes the content of polypheno-
lic compounds (Dai and Mumper, 2010; Speer and Kölling-Speer,
2006).
3.2. Characterization of the sunscreen formulations
3.2.1. Droplet size distribution
The use of a surfactant-free emulsion might be a strategy to
avoid skin irritations associated to this type of excipients (Marku
et al., 2012). In Pickering emulsions the droplet is stabilized by a
Table 3
Particle size distribution of the different solid particles proposed (mean ± SD, n = 6).

Solid Particles Particle size distribution (␮m)

3.1.1. Wettability measurements
Span d (0.1) d (0.5) d (0.9)
In the surfactant-free system stabilized by solid particles, these
will be preferentially wet by one of the liquids, with the more poorly mTiO2 37.36 ± 1.38 0.14 ± 0.01 0.19 ± 0.01 7.12 ± 0.30
wetting liquid becoming the disperse phase. Particle wettability at ZnO 19.51 ± 5.53 0.16 ± 0.01 0.57 ± 0.03 11.38 ± 3.74
ASt 1.00 ± 0.01 7.28 ± 0.01 13.52 ± 0.01 20.82 ± 0.01
the oil–water interface is quantified by the contact angle, , that
 J. Marto et al. / Industrial Crops and Products 80 (2016) 93–100
Table 4
Droplet size distribution of GCO and SCO sunscreen (mean ± SD, n = 625).
Formulation Droplet size distribution (␮m)
Span d (0.1) d (0.5) d (0.9)
GCO sunscreen 1.17 ± 0.02 3.01 ± 0.11 5.54 ± 0.28 9.49 ± 0.55
SCO sunscreen 0.90 ± 0.03 5.72 ± 0.10 8.87 ± 0.13 13.71 ± 0.34
reduction of the bare oil-water interface by adsorption of small
particles. The parameter to describe Pickering emulsions stability
is the contact angle of the adsorbed particles, as explained above.
Initially, a contact angle greater than 90◦ is required in order to
obtain a w/o emulsion. In this assay we evaluated the influence
of the oil on Pickering emulsions droplet size distribution. Both
sunscreen formulations showed a narrow droplet size distribu-
tion with a span value in the range of 0.9–1.2. The GCO sunscreen
exhibited lower mean droplet size than SCO sunscreen (Table 4).
Previous studies demonstrated that coffee extracts have extremely
strong antioxidant properties than many other food products. This
applies to both roasted and green coffee, although, the latter con-
tains even ten times higher concentration of polyphenols (Budryn
et al., 2013). Other study demonstrated that several polyphenols,
could be adsorbed into the oil–water interface, present as water-
and oil-insoluble particles and decrease the surface tension, provid-
ing very good stabilization of emulsions and, consequently, leading
a decrease in droplet size (Luo et al., 2011; Wagemaker et al., 2011).
3.2.2. Structural analysis of the sunscreen formulations
3.2.2.1. Rheology studies. The flow curves (Fig. 1(a)) showed that
the emulsions presented a shear-thinning and rheopetic behaviour.
This behaviour could influence the sunscreen performance, creat-
ing a uniform, impenetrable and protective semi-solid film over the
skin surface that is required to obtain an effective sunscreen.
In the range of 0–5 Pa, both emulsions were not disrupted
(Fig. 1(b and c). Thus, the values of G’ and G” remained linear within
this region of linear viscoelasticity, which can therefore indicate the
suitable shear stress to be used in frequency sweep and creep and
recovery tests.
The GCO sunscreen exhibited higher G’ and G” values than SCO
sunscreen. The frequency sweep curves of GCO sunscreen (Fig. 1(b))
showed that in the range tested (0.1–100 Hz) there was practically
no variation in the elastic and viscous moduli. Furthermore, vis-
coelastic behaviour was seen over the whole range, since G’ (elastic
modulus) was higher than G” (viscous modulus). The same rela-
tion is observed for SCO sunscreen, but the G’ and G” values for
GCO sunscreen are higher than for SCO sunscreen, exhibiting high
stability.
In the creep and recovery test, both sunscreens suffered defor-
mation, shown by the compliance value (J), but SCO sunscreen was
much more susceptible to this force (Fig. 1(c). In the recovery part
of this assay, when the shear stress was removed and the sam-
ples could recover their former structure, the elastic part of the
deformation, was reversed.
In summary, the addition of GCO promoted an increase in the
apparent viscosity and the elastic modulus, when compared to the
formulation prepared with SCO. Considering the results of the par-
ticle size distribution (Table 4), the reduction in droplet size results
in an increase in the viscosity and storage modulus of the emulsions
and the shear-thinning behaviour becomes even stronger, which
suggested an enhancement in emulsion stability and performance
(Pal, 1996).
3.2.2.2. TPA. The results of the calculations of textural parameters
are collected in Table 5. The adhesiveness is the only parameter,
which differs between the two formulations (p < 0.05). Concerning
the emulsions adhesiveness, which is more a surface characteristic
97
Table 5
Mechanical properties of the sunscreens extracted from the TPA mode (mean ± SD,
n = 3).
Formulations Hardness g Adhesiveness Cohesiveness Compressibility
(|g s|) (g s)
GCO sunscreen 23.62 ± 1.76 39.76 ± 0.27 0.82 ± 0.04 29.87 ± 1.91
SCO sunscreen 25.23 ± 2.26 48.65 ± 0.53 0.83 ± 0.01 31.09 ± 1.86
and depends on a combined effect of adhesive and cohesive forces,
the presence of SCO caused an increase in this parameter.
According to Alves et al. (2003), during coffee roasting process
there are changes in the fatty acid composition, increasing the
trans-fatty acid levels, which increase the hydrophobic character
of the final formulation and, consequently enhance the adhesive
forces and water repellent performance (Shyr and Ou-Yang, 2015).
Other authors proved that spent coffee oil also contains more
hydrophobic compounds, such as fatty acids, fatty acid esters,
medium-chain paraffins and olefins than green coffee oil or other
vegetable oils (Kelkar et al., 2015).
Considering the hardness, cohesiveness and compressibility
results, the lipid type did not influence these parameters.
The fatty acid tails are hydrophobic because they are non polar
and the heads are hydrophobic because they are polar.
3.2.3. In vitro SPF
Both emulsions showed high values of SPF with a suitable
UVA/UVB ratio (Table 6). For GCO there was a synergistic increase
of SPF value (around 1.6 fold) when combined with physical fil-
ters as mTiO2 in GCO sunscreen. According to Wagemaker et al.
(2011) the oil extracted from unroasted beans is used by the cos-
metics industry due to its excellent properties for the human skin,
particularly maintaining skin-moisture due to its fatty acid com-
position. In addition, its ability to absorb UV radiation in the UVB
range allows its use in sunscreens as a SPF enhancer, allowing to
reduce the amount of physical and chemical filters (Chiari et al.,
2014).
3.2.4. In vitro sun product water resistance
The in vitro method measures the SPF following a defined water
immersion procedure. For a product to be considered water resis-
tant, the value for the lower 90% one-sided confidence limit has to
be greater than or equal to 50%. A ‘waterproof’ product should have
a %WRR exceeding 80% after two immersions, while the %WRR of a
‘water resistant’ product should be more than 50% (COLIPA, 2005).
After the first immersion, the GCO sunscreen presented a %WRR
of 62.6, which decreased in the second immersion to 50.7%. In turn,
SCO sunscreen showed a value of 83.1% at first immersion and a
value of 80.8% of WRR after the second immersion. Based on this
%WRR, it is possible to ensure the ‘water resistance’ mark for GCO
sunscreen and the ‘waterproof’ claim for SCO sunscreen. As men-
tioned previously, during the coffee roasting process the fatty acid
levels increase, enhancing the hydrophobicity of the SCO sunscreen
and, consequently, improving the effectiveness of water resistance
performance (Kelkar et al., 2015). Additionally, hydrophobic sun-
screen formulations can also serve as additional water barriers to
help mitigate the disruption in stratum corneum caused by constant
exposure to water (Shyr and Ou-Yang, 2015). In water resistant
products typically more polar lipophilic ingredients can be found.
Conversely, waterproofing sunscreens should contain more non-
polar ingredients, which is the case of SCO (Couteau et al., 2012).
3.2.5. Skin adhesion properties
Table 7 shows the skin adhesion properties of the formulations.
The force necessary to detach the GCO and SCO sunscreen samples
from the skin was statistically higher than the one needed to detach
98 J. Marto et al. / Industrial Crops and Products 80 (2016) 93–100

Fig. 1. (a) Flow curves, (b) frequency sweeps and (c) creep and recovery plots of GCO sunscreen and SCO sunscreen.

the blank. In our study, when the SCO was used, the work of skin sunscreens influenced the skin adhesion ability of emulsions and,
adhesion ability after water immersion was also increased (Table 7). consequently the water resistant performance. According to the lit-
The type of lipid used in the preparation of water in oil emulsions erature (Draelos, 2011), the lipid material used in the preparation
 J. Marto et al. / Industrial Crops and Products 80 (2016) 93–100 99

Table 6
In vitro and in vivo efficacy tests of the GCO and SCO sunscreen.

Formulations In vitrosun protection factor In vitro sun product water resistance In vivosun product
(SPF)–optometrics SPF-290S Analyzer water resistance

SPF Rating UVA/UVB UVA SPF after SPF after %WRRa %WRRa after %WRRa
1st 2nd after 1st 2nd
immersion immersion immersion immersion

GCO sunscreen 82.3 ± 10.3

50+ 0.9 ± 0.1 71.1 ± 10.1 52.0 ± 14.2 42.1 ± 13.4 62.6 50.7 70.0 ± 2.2
SCO sunscreen 51.9 ± 5.550 0.9 ± 0.1 44.2 ± 5.2 43.18 ± 6.88 41.95 ± 5.94 83.1 80.8 80.0 ± 3.6
a
Water resistance retention.

Table 7
Skin adhesive properties of the formulations on dry skin and after 40 min of water immersion (mean ± SD, n = 6).

Samples Work of skin adhesion (mJ/cm2 )  of skin adhesion (mJ/cm2 )a

GCO sunscreen on dry skin 0.05 ± 0.01 –

SCO sunscreen on dry skin 0.05 ± 0.01 –
GCO sunscreen after water immersion 0.01 ± 0.01 0.04
SCO sunscreen after water immersion 0.04 ± 0.01 0.01
a
Work of skin adhesion between sunscreens on dry skin and after water immersion.

Fig. 2. Histograms of skin whiteness resulting from sunscreens applied on dry skin and cross-polarized images of two sunscreens applied to the volar forearm of a subject:
Dark grey–bare skin; grey–fresh sunscreen applications with 30 min air drying; and light grey - sunscreen after 40 min water immersion.

of w/o emulsions sunscreens influence the skin adhesion ability of
emulsions and consequently their water resistance performance.
The SCO extracted from roasted coffee is a brown viscous liquid due
to the presence of liposoluble Maillard reaction products separated
during the oil extraction, enhancing its lipophilic characteristics
(Speer and Kölling-Speer, 2006). Concerning the skin surface prop-
erties, it was demonstrated that dry skin is mostly lipophilic and
wet skin is more hydrophilic, having higher surface energy. Thus,
the surface energy values of dry skin reflect its dominant lipophilic-
ity and more apolar materials, such as SCO, may also adhere to
skin.
3.2.6. HRIPT
Under the experimental conditions adopted, the repeated appli-
cations of the sunscreens under occlusive patch induced no
irritative reaction and the product has very good skin compatibility.
Moreover, the repeated applications induced no allergic reaction.
3.2.7. In vivo sun product water resistance
Human testing is considered to be the most acceptable method
for claiming water resistance. This method is a new in vivo screening
approach to measure WRR using cross-polarized imaging. Although
it does not allow determining the exact SPF before and after the
100 J. Marto et al. / Industrial Crops and Products 80 (2016) 93–100
immersion, it allows evaluating the amount of sunscreen lost due
to the action of water.
Skin whiteness results showed a similar behaviour as obtained
for water resistance results (Fig. 2). After water immersion, both
emulsions presented a range between 50 and 80% of whiteness with
a mean peak around 70% and 75% of whiteness for GCO sunscreen
and SCO sunscreen, respectively.
GCO sunscreen and SCO sunscreen, when exposed to water for
40 min, revealed 70.0 ± 2.2% and 80.0 ± 3.6% of whiteness, respec-
tively (Table 6). In vivo WRR values confirmed the results obtained
in in vitro assay, thus supporting the “water resistant” claim of the
GCO and SCO sunscreens. Furthermore, these in vivo values were
higher than those obtained in vitro, which may be explained by a
better adhesion to human skin compared with the adhesive tape
used for the in vitro assay.
4. Conclusions
A novel sunscreen formulation with a high UVB/A protection,
biological activity and better tolerability was developed based on
the Pickering emulsions concept. The successful formulation was
possible by combining natural and multifunctional compounds.
Additionally, the coffee oils studied in this work possess all of the
properties required for sunscreens and may be considered desirable
and suitable ingredients for industrial applications. In addition to
being a green product, it contains a series of lipophilic substances
with important antioxidant characteristics, such as tocopherols,
that protect the skin against UVB radiation.
When high oil contents are combined with a composition rich
in unsaturated acids, and compounds with high SPF value and high
unsaponifiable material content, the result is a product or ingre-
dient that is ideally suited for formulating high quality cosmetic
products able to promote moisture retention and simultaneously
provide sun protection. All results revealed an excellent compro-
mise between stability, UV protection, rheological and mechanical
behaviour, efficacy, safety and cosmeticity.
The use of spent and green coffee oil in cosmetic industry seems
to be a suitable approach to recycle and valorise wastes from coffee
industry. Moreover the coffee oil presented promising characteris-
tics in the improvement of sunscreen performance.
Acknowledgments
This work was supported by the Fundação para a Ciência e a Tec-
nologia, Portugal (UID/DTP/04138/2013 to iMed.ULisboa and grant
SFRH/BDE/51599/2011), the strategic project UID/QUI/50006/2013
and Laboratórios Atral S.A., Portugal.
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